Abstract
Agrobacterium-mediated transformation of indica rice is undoubtedly a challenging task due to the rice recalcitrant nature to transformation process. Therefore, optimization of the transformation protocol is important for specific indica rice cultivar to ensure effectiveness of the transformation. In this study, crucial parameters affecting Agrobacteriummediated transformation were optimized to obtain transgenic rice of local rice cultivar (indica MR219). Embryogenic calli were chosen for inoculation with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pH2GW7-ABP57 containing gene of interest, Auxin binding protein 57 (Abp57). The parameters that have been optimized were the immersion time, co-cultivation period, acetosyringone concentration and co-cultivation temperature. A total of four days co-cultivation period and 30 min immersion of embryogenic callus are optimum for the transformation of MR219 with transformation efficiency of 26.4% and 16.0%, respectively. Acetosyringone at 200 M and co-cultivation at 28C also gave the highest transformation efficiency (14.4 and 18.4%, respectively). Meanwhile, inclusion of 20 g/L maltose+20 g/L sorbitol into the regeneration media has significantly improve the transformed somatic embryos growth and increase the regeneration efficiency up to 40.0%. The results of polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) indicated that the transgene was successfully integrated and overexpressed in transgenic rice of MR219. In conclusion, significant improvement in transformation efficiency for rice cv. MR219 has been obtained by using the optimised protocol for transformation and regeneration developed in this study.
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Claims 7
Claim · model-written summary
Four days of co-cultivation gave 26.4% transformation efficiency.
- Taxon
- Rice
Oryza sativat0 - Material
- MR219e0
- Treatment
- four days co-cultivation periode0
- Outcome
- transformation efficiencye0
- Direction
- optimume0
- Value
- 26.4%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A total of four days co-cultivation period and 30 min immersion of embryogenic callus are optimum for the transformation of MR219 with transformation efficiency of 26.4% and 16.0%, respectively.”
t0“Rice”
Claim · model-written summary
Thirty minutes of embryogenic callus immersion gave 16.0% transformation efficiency.
- Taxon
- RiceOryza sativat0
- Material
- MR219e0
- Treatment
- 30 min immersion of embryogenic calluse0
- Outcome
- transformation efficiencye0
- Direction
- optimume0
- Value
- 16.0%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A total of four days co-cultivation period and 30 min immersion of embryogenic callus are optimum for the transformation of MR219 with transformation efficiency of 26.4% and 16.0%, respectively.”
t0“Rice”
Claim · model-written summary
Acetosyringone at 200 M gave the highest transformation efficiency, 14.4%.
- Taxon
- RiceOryza sativat0
- Treatment
- Acetosyringone at 200 Me0
- Outcome
- transformation efficiencye0
- Direction
- higheste0
- Value
- 14.4e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Acetosyringone at 200 M and co-cultivation at 28C also gave the highest transformation efficiency (14.4 and 18.4%, respectively).”
t0“Rice”
Claim · model-written summary
Co-cultivation at 28C gave the highest transformation efficiency, 18.4%.
- Taxon
- RiceOryza sativat0
- Treatment
- co-cultivation at 28Ce0
- Outcome
- transformation efficiencye0
- Direction
- higheste0
- Value
- 18.4%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Acetosyringone at 200 M and co-cultivation at 28C also gave the highest transformation efficiency (14.4 and 18.4%, respectively).”
t0“Rice”
Claim · model-written summary
Maltose and sorbitol improved growth of transformed somatic embryos.
- Taxon
- RiceOryza sativat0
- Treatment
- 20 g/L maltose+20 g/L sorbitole0
- Outcome
- transformed somatic embryos growthe0
- Direction
- improvee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Meanwhile, inclusion of 20 g/L maltose+20 g/L sorbitol into the regeneration media has significantly improve the transformed somatic embryos growth and increase the regeneration efficiency up to 40.0%.”
t0“Rice”
Claim · model-written summary
Maltose and sorbitol increased regeneration efficiency to 40.0%.
- Taxon
- RiceOryza sativat0
- Treatment
- 20 g/L maltose+20 g/L sorbitole0
- Outcome
- regeneration efficiencye0
- Direction
- increasee0
- Value
- 40.0%e0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Meanwhile, inclusion of 20 g/L maltose+20 g/L sorbitol into the regeneration media has significantly improve the transformed somatic embryos growth and increase the regeneration efficiency up to 40.0%.”
t0“Rice”
Claim · model-written summary
PCR assays indicated successful integration and overexpression of the transgene in MR219 rice.
- Taxon
- riceOryza sativae0
- Material
- MR219e0
- Gene
- Abp57t0
- Outcome
- transgene was successfully integrated and overexpressede0
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The results of polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) indicated that the transgene was successfully integrated and overexpressed in transgenic rice of MR219.”
t0“Abp57 Gene”
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